饥饿对Beclin-1依赖的Ana-1细胞自噬和凋亡的影响

doi:10.13481/j.1671-587x.20180403

吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (04): 704-708.doi: 10.13481/j.1671-587x.20180403

饥饿对Beclin-1依赖的Ana-1细胞自噬和凋亡的影响

杨万霞^1,2, 苏强³, 邢爱华², 张晓延^2,4

1. 兰州大学第二医院检验科, 甘肃兰州 730000;
2. 山西医科大学汾阳学院医学检验系, 山西汾阳 032200;
3. 山西医科大学第一医院神经内科, 山西太原 030001;
4. 山西医科大学药学院, 山西太原 030001

收稿日期:2017-10-31 出版日期:2018-07-28 发布日期:2018-07-27
通讯作者: 张晓延,副教授,硕士研究生导师(Tel:0358-2100627,E-mail:fyzxy@sohu.com) E-mail:fyzxy@sohu.com
作者简介:杨万霞(1992-),女,甘肃省陇南市人,在读医学硕士,主要从事临床检验诊断学方面的研究。
基金资助:
国家自然科学基金资助课题（81301426）；山西省教育厅高等院校大学生创新创业项目资助课题（2016526）

Effects of starvation on autophagy and apoptosis of Beclin-1-dependent Ana-1 cells

YANG Wanxia^1,2, SU Qiang³, XING Aihua², ZHANG Xiaoyan^2,4

1. Clinical Laboratory, Second Hospital, Lanzhou University, Lanzhou 730000, China;
2. Department of Medical Laboratory, Fenyang College, Shanxi Medical University, Fenyang 032200, China;
3. Department of Neurology, First Hospital, Shanxi Medical University, Taiyuan 030001, China;
4. College of Pharmacy, Shanxi Medical University, Taiyuan 030001, China

Received:2017-10-31 Online:2018-07-28 Published:2018-07-27

摘要/Abstract

摘要： 目的：研究饥饿对Ana-1细胞自噬和凋亡的影响，并探讨其作用机制。方法：体外培养Ana-1细胞，将细胞分为对照组（饥饿0h）和饥饿3、6、9、12、24h组（饥饿组），观察饥饿对细胞自噬与凋亡的影响；将Ana-1细胞分为空白对照组、饥饿组、3-甲基腺嘌呤（3-MA）组、3-MA联合饥饿组，孵育24 h，采用Western blotting法检测各组细胞中自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ比值及Beclin-1蛋白表达水平和凋亡相关蛋白Caspase-3蛋白水平，采用倒置显微镜观察饥饿6、12和24 h组细胞形态表现。结果：与对照组比较，饥饿6、12和24h组Ana-1细胞中LC3-Ⅱ/LC3-Ⅰ比值，饥饿3、6、9、12和24 h组Ana-1细胞中Beclin-1蛋白表达水平均明显升高（P<0.05或P<0.01），饥饿3、6、9、12和24 h组Ana-1细胞中Caspase-3水平明显降低（P<0.01），呈时间依赖性；倒置显微镜下观察，与对照组比较，不同时间饥饿组细胞形态发生改变，细胞皱缩、出现碎片，随着时间的延长，细胞变化更加明显。与饥饿组比较，3-MA联合饥饿组Ana-1细胞中LC3-Ⅱ/LC3-Ⅰ比值和Beclin-1蛋白表达水平及Caspase-3蛋白水平均明显降低（P<0.05或P<0.01）。结论：饥饿可诱导Beclin-1依赖的Ana-1细胞自噬并引起细胞凋亡。

关键词: 细胞凋亡, 饥饿, Beclin-1, Ana-1细胞, 细胞自噬

Abstract: Objective:To study the effects of starvation on the autophagy and apoptosis of Ana-1 cells,and to explore their mechanisms. Methods:The Ana-1 cells were cultured in vitro and starved for 0 h(control group),3,6,9,12 and 24 h(starvation group)to study the effects of starvation on the autophagy and apoptosis.The Ana-1 cells were divided into blank control group,starvation group,3-methyladenine group(3-MA) and 3-MAcombined with starvation group,and they were incubated for 24 h to explore their mechanisms.The ratio of autophagy associated protein LC3-Ⅱ/LC3-Ⅰ,the expression levels of Beclin-1 and apoptosis-related protein Caspase-3 in the cells in various groups were detected by Western blotting method.Cell morphology 6,12,and 24 h after starvation was observed by inverted microscope. Results:Compared with control group,the ratio of LC3-Ⅱ/LC3-Ⅰ in the Ana-1 cells in starvation group(6,12,and 24 h)and the expression levels of Beclin-1 in starvation group(3,6,9,12,and 24 h) were significantly increased(P<0.05 or P<0.01),and the Caspase-3 levels in starvation group(3,6,9,12,and 24 h) were decreased significantly(P<0.05 or P<0.01) in a time-dependent manner.Compared with control group,the cells in starvation group at different time were changed in shape:the cells shrank,the fragments appeared,and the changes of cells became more obvious with the prolongation of time.Compared with starvation group,the ratio of LC3-Ⅱ/LC3-Ⅰ and the expression levels of Beclin-1 and Caspase-3 in the Ana-1 cells in 3-MA combined with starvation group were decreased significantly(P<0.05 or P<0.01). Conclusion:Starvation could induce the autophagy and apoptosis of the Beclin-1-dependent Ana-1 cells.

Key words: Beclin-1, Ana-1 cells, cell autophagy, apoptosis, starvation

中图分类号:

Q255

杨万霞, 苏强, 邢爱华, 张晓延. 饥饿对Beclin-1依赖的Ana-1细胞自噬和凋亡的影响[J]. 吉林大学学报(医学版), 2018, 44(04): 704-708.

YANG Wanxia, SU Qiang, XING Aihua, ZHANG Xiaoyan. Effects of starvation on autophagy and apoptosis of Beclin-1-dependent Ana-1 cells[J]. Journal of Jilin University Medicine Edition, 2018, 44(04): 704-708.

参考文献

[1] Tsujimoto Y,Shimizu S.Another way to die:autophagic programmed cell death[J].Cell Death Differ,2005,12(Suppl2):1528-1534.
[2] Klionsky DJ,Emr SD.Autophagy as a regulated pathway of cellular degradation[J].Science,2000,290(5497):1717-1721.
[3] Kim SY,Song X,Zhang L,et al.Role of Bcl-xL/Beclin-1 in interplay between apoptosis and autophagy in oxaliplatin and bortezomib-induced cell death[J].Biochem Pharmacol,2014,88(2):178-188.
[4] Yang J,Yao S.JNK-Bcl-2/Bcl-xL-Bax/Bak pathway mediates the crosstalk between matrine-induced autophagy and apoptosis via interplay with Beclin 1[J].Int J Mol Sci,2015,16(10):25744-25758.
[5] Shintani T,Klionsky DJ.Autophagy in health and disease:a double-edged sword[J].Science,2004,306(5698):990-995.
[6] Mans LA,Querol Cano L,van Pelt J,et al.The tumor suppressor LKB1 regulates starvation-induced autophagy under systemic metabolic stress[J].Sci Rep,2017,7(1):7327.
[7] Piano Mortari E,Folgiero V,Marcellini V,et al.The Vici syndrome protein EPG5 regulates intracellular nucleic acid trafficking linking autophagy to innate and adaptive immunity[J].Autophagy,2018,14(1):22-37.
[8] Kihara A,Kabeya Y,Ohsumi Y,et al.Beclin-phosphatidylinositol 3-kinase complex functions at the trans-Golgi network[J].EMBO Rep,2001,2(4):330-335.
[9] Liang XH,Jackson S,Seaman M,et al.Induction of autophagy and inhibition of tumorigenesis by beclin 1[J].Nature,1999,402(6762):672-676.
[10] Liang XH,Kleeman LK,Jiang HH,et al.Protection against fatal Sindbis virus encephalitis by beclin,a novel Bcl-2-interacting protein[J].J Virol,1998,72(11):8586-8596.
[11] McKnight NC,Zhenyu Y.Beclin 1,an essential component and master regulator of PI3K-Ⅲ in health and disease[J].Curr Pathobiol Rep,2013,1(4):231-238.
[12] McKnight NC,Zhong Y,Wold MS,et al.Beclin 1 is required for neuron viability and regulates endosome pathways via the UVRAG-VPS34 complex[J].PLoS Genet,2014,10(10):e1004626.
[13] 安洋,张慧宇,郭俊峰,等.降钙素基因相关肽对血清饥饿作用下MC3T3-E1成骨细胞凋亡和自噬的影响[J].华西口腔医学杂志,2017,35(2):133-138.
[14] Wu YT,Tan HL,Shui G,et al.Dual role of 3-methyladenine in modulation of autophagy via different temporal patterns of inhibition on class Ⅰ and Ⅱ phosphoinositide 3- kinase[J].J Biol Chem,2010,285(14):10850-10861.
[15] Huang Y,Fu Z,Dong W,et al.Serum starvation-induces down-regulation of Bcl-2/Bax confers apoptosis in tongue coating-related cells in vitro[J].Mol Med Rep,2018,17(4):5057-5064.
[16] Gaytan M,Morales C,Sánchez-Criado JE,et al.Immunolocalization of beclin 1,a bcl-2-binding,autophagy-related protein,in the human ovary:possible relation to life span of corpus luteum[J].Cell Tissue Res,2008,331(2):509-517.
[17] Qin B,Zhou Z,He J,et al.IL-6 inhibits starvation-induced autophagy via the STAT3/Bcl-2 signaling pathway[J].Sci Rep,2015,5:15701.
[18] Zhong L,Shu W,Dai W,et al.Reactive oxygen species-mediated c-Jun NH2-terminal kinase activation contributes to hepatitis B virus X protein-induced autophagy via regulation of the Beclin-1/Bcl-2 interaction[J].J Virol,2017,91(15):e00001-e00017.
[19] 杨翼,李章华,贺磊磊,等.自噬抑制剂3-MA对"饥饿状态"下心肌细胞自噬和凋亡的影响[J].武汉体育学院学报,2014,48(11):61-66.
[20] 王晓娜,任来峰,赵安江,等.3-甲基腺嘌呤对喜树碱诱导的宫颈癌Hela细胞凋亡的影响[J].中国免疫学杂志,2016,32(8):1128-1132.
[21] Wirawan E,Vande Walle L,Kersse K,et al.Caspase-mediated cleavage of Beclin-1 inactivates Beclin-1-induced autophagy and enhances apoptosis by promoting the release of proapoptotic factors from mitochondria[J].Cell Death Dis,2010,1:e18.
[22] Zhang BC,Zhang CW,Wang C,et al.Luteolin attenuates foam cell formation and apoptosis in ox-LDL-stimulated macrophages by enhancing autophagy[J].Cell Physiol Biochem,2016,39(5):2065-2076.

饥饿对Beclin-1依赖的Ana-1细胞自噬和凋亡的影响

Effects of starvation on autophagy and apoptosis of Beclin-1-dependent Ana-1 cells

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